Abstract
MicroRNAs are tiny molecules that strikingly change their expression patterns and distribution during somatic embryogenesis induction and plant regeneration. It is of great relevance to analyze simultaneously the microRNA and target mRNA fates to understand their role in promoting an adequate embryogenic response to external stimulus in the regenerating tissues. Here we describe a method to evaluate the expression patterns of miRNAs or other sRNAs and their target regulation in distinctive tissues observed during maize plant regeneration. Key features of the method include the classification of regenerating plant material with reproducibly distinctive morphological characteristics and a purification procedure that renders high-quality small and large RNA separation from the same sample for qRT-PCR analysis.
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Acknowledgment
Research performed in Dr. Dinkova’s lab is supported by grants from Consejo Nacional de Ciencia y Tecnología, 238439, Programa de Apoyo a Proyectos de Investigación e Innovación Tecnológica, IN211215, IN214118, and PAIP 5000-9118. The authors appreciate the technical assistance provided by Maria Teresa de Jesús Olivera Flores during in vitro plant tissue culture.
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López-Ruiz, B.A., Juárez-González, V.T., Chávez-Hernández, E.C., Dinkova, T.D. (2018). MicroRNA Expression and Regulation During Maize Somatic Embryogenesis. In: Loyola-Vargas, V., Ochoa-Alejo, N. (eds) Plant Cell Culture Protocols. Methods in Molecular Biology, vol 1815. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8594-4_28
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DOI: https://doi.org/10.1007/978-1-4939-8594-4_28
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